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The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor
We previously identified miR-4731-5p (miR-4731) as a melanoma-enriched microRNA following comparison of melanoma with other cell lines from solid malignancies. Additionally, miR-4731 has been found in serum from melanoma patients and expressed less abundantly in metastatic melanoma tissues from stag...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226538/ https://www.ncbi.nlm.nih.gov/pubmed/27331623 http://dx.doi.org/10.18632/oncotarget.10109 |
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author | Stark, Mitchell S. Tom, Lisa N. Boyle, Glen M. Bonazzi, Vanessa F. Soyer, H. Peter Herington, Adrian C. Pollock, Pamela M. Hayward, Nicholas K. |
author_facet | Stark, Mitchell S. Tom, Lisa N. Boyle, Glen M. Bonazzi, Vanessa F. Soyer, H. Peter Herington, Adrian C. Pollock, Pamela M. Hayward, Nicholas K. |
author_sort | Stark, Mitchell S. |
collection | PubMed |
description | We previously identified miR-4731-5p (miR-4731) as a melanoma-enriched microRNA following comparison of melanoma with other cell lines from solid malignancies. Additionally, miR-4731 has been found in serum from melanoma patients and expressed less abundantly in metastatic melanoma tissues from stage IV patients relative to stage III patients. As miR-4731 has no known function, we used biotin-labelled miRNA duplex pull-down to identify binding targets of miR-4731 in three melanoma cell lines (HT144, MM96L and MM253). Using the miRanda miRNA binding algorithm, all pulled-down transcripts common to the three cell lines (n=1092) had potential to be targets of miR-4731 and gene-set enrichment analysis of these (via STRING v9.1) highlighted significantly associated genes related to the ‘cell cycle’ pathway and the ‘melanosome’. Following miR-4731 overexpression, a selection (n=81) of pull-down transcripts underwent validation using a custom qRT-PCR array. These data revealed that miR-4731 regulates multiple genes associated with the cell cycle (e.g. CCNA2, ORC5L, and PCNA) and the melanosome (e.g. RAB7A, CTSD, and GNA13). Furthermore, members of the synovial sarcoma X breakpoint family (SSX) (melanoma growth promoters) were also down-regulated (e.g. SSX2, SSX4, and SSX4B) as a result of miR-4731 overexpression. Moreover, this down-regulation of mRNA expression resulted in ablation or reduction of SSX4 protein, which, in keeping with previous studies, resulted in loss of 2D colony formation. We therefore speculate that loss of miR-4731 expression in stage IV patient tumours supports melanoma growth by, in part; reducing its regulatory control of SSX expression levels. |
format | Online Article Text |
id | pubmed-5226538 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-52265382017-01-18 The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor Stark, Mitchell S. Tom, Lisa N. Boyle, Glen M. Bonazzi, Vanessa F. Soyer, H. Peter Herington, Adrian C. Pollock, Pamela M. Hayward, Nicholas K. Oncotarget Research Paper We previously identified miR-4731-5p (miR-4731) as a melanoma-enriched microRNA following comparison of melanoma with other cell lines from solid malignancies. Additionally, miR-4731 has been found in serum from melanoma patients and expressed less abundantly in metastatic melanoma tissues from stage IV patients relative to stage III patients. As miR-4731 has no known function, we used biotin-labelled miRNA duplex pull-down to identify binding targets of miR-4731 in three melanoma cell lines (HT144, MM96L and MM253). Using the miRanda miRNA binding algorithm, all pulled-down transcripts common to the three cell lines (n=1092) had potential to be targets of miR-4731 and gene-set enrichment analysis of these (via STRING v9.1) highlighted significantly associated genes related to the ‘cell cycle’ pathway and the ‘melanosome’. Following miR-4731 overexpression, a selection (n=81) of pull-down transcripts underwent validation using a custom qRT-PCR array. These data revealed that miR-4731 regulates multiple genes associated with the cell cycle (e.g. CCNA2, ORC5L, and PCNA) and the melanosome (e.g. RAB7A, CTSD, and GNA13). Furthermore, members of the synovial sarcoma X breakpoint family (SSX) (melanoma growth promoters) were also down-regulated (e.g. SSX2, SSX4, and SSX4B) as a result of miR-4731 overexpression. Moreover, this down-regulation of mRNA expression resulted in ablation or reduction of SSX4 protein, which, in keeping with previous studies, resulted in loss of 2D colony formation. We therefore speculate that loss of miR-4731 expression in stage IV patient tumours supports melanoma growth by, in part; reducing its regulatory control of SSX expression levels. Impact Journals LLC 2016-06-16 /pmc/articles/PMC5226538/ /pubmed/27331623 http://dx.doi.org/10.18632/oncotarget.10109 Text en Copyright: © 2016 Stark et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Paper Stark, Mitchell S. Tom, Lisa N. Boyle, Glen M. Bonazzi, Vanessa F. Soyer, H. Peter Herington, Adrian C. Pollock, Pamela M. Hayward, Nicholas K. The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor |
title | The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor |
title_full | The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor |
title_fullStr | The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor |
title_full_unstemmed | The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor |
title_short | The ‘melanoma-enriched’ microRNA miR-4731-5p acts as a tumour suppressor |
title_sort | ‘melanoma-enriched’ microrna mir-4731-5p acts as a tumour suppressor |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226538/ https://www.ncbi.nlm.nih.gov/pubmed/27331623 http://dx.doi.org/10.18632/oncotarget.10109 |
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